Skin Care Composition and Method of Using the Same

ABSTRACT

A skin care composition that contains a combination of a vitamin B 3  compound, palmitoyl pentapeptide-4, acetyl tetrapeptide-11, and a dermatologically acceptable carrier. The combination of vitamin B 3  compound, palmitoyl pentapeptide-4, and acetyl tetrapeptide-11 increases activation of a cell&#39;s Antioxidant Response Element (ARE), (e.g., synergistically) to improve a visible sign of skin aging.

FIELD

The present disclosure is directed generally to improving skin healthwith a synergistic combination of skin care actives. More specifically,the present disclosure is directed to a combination of vitamin B₃ andtwo or more peptides that synergistically stimulates cellularanti-oxidancy and repair processes.

BACKGROUND

Skin is the first line of defense against environmental insults thatwould otherwise damage sensitive underlying tissue and organs, and skinplays a key role in a person's physical appearance. The tell-tale signsof aging, such as wrinkles and age spots on the skin, are an undesirablereminder of the disappearance of youth. As a result, treating the signsof aging in skin has become a booming business in youth-conscioussocieties.

Skin is made up of a variety of different cells that function togetherin a dynamic, complex relationship to maintain the health of the skin.However, as skin cells age or become damaged, they generally lose theirability to function at the level needed to maintain young, healthylooking skin. Skin cells can be damaged by a variety of endogenous andexogenous stressors (e.g., ultraviolet radiation, pollution, smoking).In some instances, these stressors cause the production of reactiveoxygen species (ROS), which interfere with normal cellular processes. Inresponse, human cells have evolved defenses to combat ROS, but thecell's defenses can be overwhelmed by spikes of stressor-induced ROS,leading to not just acute but also chronic alterations in cellularhomeostasis. As ROS accumulate over time, they cause oxidative stress atthe cellular level, which can ultimately manifest as visible signs ofaging (e.g., fine lines, wrinkles, hyperpigmented spots, thinning skin).

The antioxidant defense system of most cells is typically controlled bya master switch referred to as the antioxidant response element (ARE).The ARE is a cis-acting enhancer sequence located in the regulatoryregions of antioxidant and detoxifying genes. The ARE is activated byredox-cycling phenols and electrophiles. Some studies have shown that aprotein called nuclear factor erythroid 2-related factor 2 (Nrf2) may bethe principal transcription factor necessary for ARE activation, eventhough many other transcription factors also bind to the ARE sequence.When the ARE is activated in response to damage from ROS, thecorresponding genes signal the cell to begin producingreduction/oxidation regulators and/or ROS quenching proteins andenzymes. In addition to modulating the production of redox regulatorsand ROS quenching compounds, the ARE can also signal the cell to beginrepair processes. Thus, it would be desirable to boost activation of theARE in aging skin cells to combat the signs of skin aging caused byoxidative stress.

A number of compounds have been discovered that boost activation of theARE. For example, US20160317419 provides data showing that a combinationof niacinamide and nicotinamide riboside boosts ARE activation. However,there is still a need to identify compounds and methods that improvecellular anti-oxidancy by boosting ARE activation. There is also a needto identify compounds and methods that beneficially modulate genesinvolved in cellular anti-oxidancy and repair.

SUMMARY

The present disclosure relates to a skin care composition, comprising acombination of a vitamin B₃ compound, palmitoyl pentapeptide-4(pal-KTTKS) [SEQ ID NO: 1], acetyl tetrapeptide-11 (ac-PPYL) [SEQ ID NO:2], and a dermatologically acceptable carrier. The combination ofvitamin B₃ compound, pal-KTTKS [SEQ ID NO: 1], and ac-PPYL [SEQ ID NO:2]. This combination of compounds has been found to activate a cell'sAntioxidant Response Element (ARE) and/or beneficially modulate genesinvolved in cellular anti-oxidancy and repair, in some instancessynergistically. The present disclosure also relates to methods of usingthe foregoing compositions.

DETAILED DESCRIPTION

Improving cellular anti-oxidancy and repair is important for improvingthe health and/or appearance of skin. Thus, there is a long felt need toidentify new ingredients for use in topical skin care compositions thatprovide these benefits. In particular, there is a need to identifycompounds that boost ARE activation in skin cells, which tend to be moreat-risk from oxidative stress than some other types of cells due totheir high exposure to ultraviolet radiation and other exogenousstressors. It has now been surprisingly discovered that combining avitamin B₃ compound, palmitoyl pentapeptide-4 [SEQ ID NO: 1], and acetyltetrapeptide-11 [SEQ ID NO: 2] can stimulate anti-oxidancy and cellrepair processes. In particular, this combination of ingredients hasbeen found to synergistically boost ARE activation and/or modulatecertain genes believed to be involved in cellular anti-oxidancy andrepair.

Reference herein to “embodiment(s)” or the like means that a particularmaterial, feature, structure and/or characteristic described inconnection with the embodiment is included in at least one embodiment,optionally a number of embodiments, but it does not mean that allembodiments incorporate the material, feature, structure, and/orcharacteristic described. Furthermore, materials, features, structuresand/or characteristics may be combined in any suitable manner acrossdifferent embodiments, and materials, features, structures and/orcharacteristics may be omitted or substituted from what is described.Thus, embodiments and aspects described herein may comprise or becombinable with elements or components of other embodiments and/oraspects despite not being expressly exemplified in combination, unlessotherwise stated or an incompatibility is stated.

In all embodiments, all ingredient percentages are based on the weightof the cosmetic composition, unless specifically stated otherwise. Allratios are weight ratios, unless specifically stated otherwise. Thenumber of significant digits conveys neither a limitation on theindicated amounts nor on the accuracy of the measurements. All numericalamounts are understood to be modified by the word “about” unlessotherwise specifically indicated. Unless otherwise indicated, allmeasurements are understood to be made at approximately 25° C. and atambient conditions, where “ambient conditions” means conditions underabout 1 atmosphere of pressure and at about 50% relative humidity. Allnumeric ranges are inclusive and combinable to form narrower ranges notexplicitly disclosed. For example, delineated upper and lower rangelimits are interchangeable to create further ranges.

The compositions of the present invention can comprise, consistessentially of, or consist of, the essential components as well asoptional ingredients described herein. As used herein, “consistingessentially of” means that the composition or component may only includeadditional ingredients that do not materially alter the basic and novelcharacteristics of the claimed composition or method. As used in thedescription and the appended claims, the singular forms “a”, “an”, and“the” are intended to include the plural forms as well, unless thecontext clearly indicates otherwise.

Sequence Listing

A sequence listing that sets forth the amino acid sequences for SEQ IDNOS: 1 and 2 and the nucleotide sequences for SEQ ID NOS: 3 to 7, whichare primary sequences and include conservatively modified variantsthereof, is being filed concurrently with the present application as anASCII text file titled “15964-Sequence_Listing_2023_07_25”. This ASCIItext file was created on Jul. 25, 2023 and is 202 KB in size. An ASCIItext file titled “15964_seq_list_ST25” was created on Jan. 19, 2021 andis 201 KB in size and was filed with U.S. patent application Ser. No.17/155,327, which this application claims benefit to. In accordance withMPEP § 605.08 and 37 CFR § 1.52(e), the subject matter in the ASCII textfiles are incorporated herein by reference.

Definitions

“About” modifies a particular value by referring to a range equal toplus or minus twenty percent (+/−20%) or less (e.g., less than 15%, 10%,or even less than 5%) of the stated value.

“Apply” or “application,” as used in reference to a composition ormaterial herein, means to place or spread the composition onto a humanskin surface such as the epidermis.

“Cosmetic composition” means a composition comprising a cosmetic agentand intended for non-therapeutic (i.e., medical) use. Examples ofcosmetic compositions include color cosmetics (e.g., foundations,lipsticks, concealers, and mascaras), skin care compositions (e.g.,moisturizers and sunscreens), personal care compositions (e.g.,rinse-off and leave on body washes and soaps), hair care compositions(e.g., shampoos and conditioners).

“Derivative” means an amide, ether, ester, amino, carboxyl, acetyl, oralcohol derivative of a given compound.

“Effective amount” means an amount of a compound or compositionsufficient to significantly induce a positive benefit to keratinoustissue over the course of a treatment period.

The positive benefit may be a health, appearance, and/or feel benefit,including, independently or in combination, the benefits disclosedherein.

“Modulate” and variations thereof mean upregulating or downregulatinggene expression.

“Skin care” means regulating and/or improving a skin condition (e.g.,skin health, appearance, or texture/feel). Some nonlimiting examples ofimproving a skin condition include improving skin appearance and/or feelby providing a smoother, more even appearance and/or feel; increasingthe thickness of one or more layers of the skin; improving theelasticity or resiliency of the skin; improving the firmness of theskin; and reducing the oily, shiny, and/or dull appearance of skin,improving the hydration status or moisturization of the skin, improvingthe appearance of fine lines and/or wrinkles, improving skin exfoliationor desquamation, plumping the skin, improving skin barrier properties,improve skin tone, reducing the appearance of redness or skin blotches,and/or improving the brightness, radiancy, or translucency of skin.

“Skin care active” means a compound or combination of compounds that,when applied to skin, provide an acute and/or chronic benefit to skin ora type of cell commonly found therein. Skin care actives may regulateand/or improve skin or its associated cells (e.g., improve skinelasticity, hydration, skin barrier function, and/or cell metabolism).

“Skin care composition” means a composition that includes a skin careactive and regulates and/or improves skin condition.

“Skin cell” refers to the types of cells commonly found in human skin.Non-limiting examples of skin cells are keratinocytes, fibroblasts,melanocytes, Langerhans cells, and Merkel cells.

“Synergy” and variations thereof mean that the cellular anti-oxidancyand repair effect provided by a combination of niacinamide, palmitoylpentapeptide-4 [SEQ ID NO: 1], and acetyl tetrapeptide-11 [SEQ ID NO: 2]is more than the predicted additive effect of these ingredients alone.For example, synergy is demonstrated when the combination ofniacinamide, palmitoyl pentapeptide-4 [SEQ ID NO: 1], and acetyltetrapeptide-11 [SEQ ID NO: 2] increases ARE activation by a more thanthe calculated additive effects of these three ingredients individually.ARE activation level can be quantitated using the ARE Assay described inmore detail below.

“Treatment period,” as used herein, means the length of time and/orfrequency that a material or composition is applied to a target skinsurface.

“Upregulation” and variations thereof mean increasing gene expression.Conversely, “downregulation” and variations thereof mean decreasing geneexpression. Gene expression can be quantitated using conventionalmethods (e.g., microarray analysis, rt-PCR, Western blot).

Skin Care Composition

The novel skin care compositions herein are intended for topicalapplication to human skin to provide a cellular anti-oxidancy and/orrepair benefit. The present skin care compositions contain a safe andeffective amount of a vitamin B3 compound, palmitoyl pentapeptide-4(pal-KTTKS) [SEQ ID NO: 1], and acetyl tetrapeptide-11 (ac-PPYL) [SEQ IDNO: 2]. An effective amount of these three ingredients in combinationcan synergistically boost activation of the antioxidant responseelement, which is important for combating oxidative stress and reducingthe visible signs of skin aging.

The combination of vitamin B3 compound, pal-KTTKS [SEQ ID NO: 1], andac-PPYL [SEQ ID NO: 2] may also synergistically modulate one or moregenes selected from Nuclear Factor Erythroid2-Related Factor 2 (NRF2)[SEQ ID NO: 3] (the primary sequence) and conservatively modifiedvariants thereof, Schlafen Family Member 5 (SLFNS) [SEQ ID NO: 4] (theprimary sequence) and conservatively modified variants thereof,Glycerophosphodiester Phosphodiesterase 1 (GDE1) [SEQ ID NO: 5] (theprimary sequence) and conservatively modified variants thereof, MultipleInositol-Polyphosphate Phosphatase 1 (MINPP1) [SEQ ID NO: 6] (theprimary sequence) and conservatively modified variants thereof, and3-Hydroxy-3-Methylglutaryl-CoA Lyase (HMGCL) [SEQ ID NO: 7] (the primarysequence) and conservatively modified variants thereof. It is believed,without being limited by theory, that these genes play important rolesin cellular anti-oxidancy and/or repair, and it has been shown thatthese genes are downregulated as a result of chronological aging and/orphotoaging in the epidermis and/or dermis. For example, NRF2 is believedto play an important role in activating the ARE (Annu Rev PharmacolToxicol. 2013; 53: 401-426). In another example, it is believed thatSLFNS plays a role in controlling extracellular matrix (ECM) remodelingenzymes. Conditions of oxidative stress stimulate ECM degradation byup-regulating metalloproteinases (MMPs). Up-regulation of SLFNS turnsdown the expression of these MMPs thus enhancing ECM integrity.

With respect to boosting ARE activation and/or modulating the expressionof one or more of the genes described above, the combination of vitaminB₃ compound, pal-KTTKS [SEQ ID NO: 1], and ac-PPYL [SEQ ID NO: 2] mayexhibit a synergy of factor of 1.2 or more (e.g., greater than 1.3, 1.4,1.5, 1.6, 1.7, 1.8, 1.9, or even 2.0) relative to the sum of theresponses from niacinamide, pal-KTTKS, and ac-PPYL treatmentsindividually, such as a vehicle control or predetermined thresholdvalue. In some instances, the composition may contain a weight ratio ofvitamin B3 compound to pal-KTTKS [SEQ ID NO: 1] to ac-PPYL [SEQ ID NO:2] of between 500:1:2 and 1:1:0.5 (e.g., between 50:1:2 and 1:1:1). Amethod for determining synergy factor is described in more detail below.

The skin care compositions herein may be cosmetic compositions,pharmaceutical compositions, or cosmeceutical compositions, and may beprovided in various product forms, including, but not limited to,solutions, suspensions, lotions, creams, gels, toners, sticks, sprays,aerosols, ointments, cleansing liquid washes and solid bars, pastes,foams, mousses, shaving creams, wipes, strips, patches,electrically-powered patches, hydrogels, film-forming products, facialand skin masks (with and without insoluble sheet), make-up such asfoundations, eye liners, and eye shadows, and the like. In someinstances, the composition form may follow from the particulardermatologically acceptable carrier chosen. For example, the composition(and carrier) may be provided in the form of an emulsion (e.g.,water-in-oil, oil-in-water, or water-in-oil-in water) or an aqueousdispersion.

The compositions herein may be prepared by conventional methods ofmaking topical skin care compositions. Such methods typically involvemixing of the ingredients in one or more steps to a relatively uniformstate, with or without heating, cooling, application of vacuum, and thelike. The compositions are preferably prepared such as to optimizestability (physical stability, chemical stability, photostability)and/or delivery of the active materials. This optimization may includeappropriate pH (e.g., less than 7), exclusion of materials that cancomplex with the active agent and thus negatively impact stability ordelivery (e.g., exclusion of contaminating iron), use of approaches toprevent complex formation (e.g., appropriate dispersing agents or dualcompartment packaging), use of appropriate photostability approaches(e.g., incorporation of sunscreen/sunblock, use of opaque packaging),etc.

Vitamin B₃ Compound

The compositions herein include a safe and effective amount of a vitaminB₃ compound. In some instances, the present compositions may contain0.01% to 10%, by weight, of the vitamin B₃ compound, based on the weightor volume of the composition (e.g., 0.1% to 10%, 0.5% to 5%, or even 1%to 3%).

As used herein, “vitamin B₃ compound” means a compound having theformula:

Where: R is CONH₂ (i.e., niacinamide), COOH (i.e., nicotinic acid) orCH₂OH (i.e., nicotinyl alcohol); derivatives thereof; and salts of anyof the foregoing.

Exemplary derivatives of vitamin B₃ compounds include nicotinic acidesters, including non-vasodilating esters of nicotinic acid (e.g.,tocopheryl nicotinate, myristyl nicotinate) nicotinamide riboside,nicotinyl amino acids, nicotinyl alcohol esters of carboxylic acids,nicotinic acid N-oxide, and niacinamide N-oxide.

Pentapeptide

The compositions herein include a safe and effective amount of thepalmitoylated pentapeptide, pal-KTTKS [SEQ ID NO: 1] (INCI: PalmitoylPentapeptide-4). In some instances, pal-KTTKS may be present in thepresent compositions at 0.0001% to 3% (e.g., 0.001% to 2%, to 1% or 0.1%to 0.5%). Pal-KTTKS is available as PROMATRIXYL from Sederma (France).

Tetrapeptide

The compositions herein include a safe and effective amount of theacetylated tetrapeptide, ac-PPYL [SEQ ID NO: 2] (INCI: AcetylTetrapeptide-11). In some instances, ac-PPYL may be present in thepresent composition at 0.0001% to 3% (e.g., 0.001% to 2%, 0.01% to 1% or0.1% to 0.5%). Ac-PPYL is available as SYNIORAGE from BASF CareCreations (New Jersey).

Dermatologically Acceptable Carrier

The compositions herein include a dermatologically acceptable carrier(which may be referred to as a “carrier”). The phrase “dermatologicallyacceptable carrier” means that the carrier is suitable for topicalapplication to the keratinous tissue, has good aesthetic properties, iscompatible with the actives in the composition, and will not cause anyunreasonable safety or toxicity concerns. In one embodiment, the carrieris present at a level of from about 50% to about 99%, about 60% to about98%, about 70% to about 98%, or, alternatively, from about 80% to about95%, by weight of the composition.

The carrier can be in a wide variety of forms. In some instances, thesolubility or dispersibility of the components (e.g., extracts,sunscreen active, additional components) may dictate the form andcharacter of the carrier. Non-limiting examples include simple solutions(e.g., aqueous or anhydrous), dispersions, emulsions, and solid forms(e.g., gels, sticks, flowable solids, or amorphous materials). In someinstances, the dermatologically acceptable carrier is in the form of anemulsion that has a continuous aqueous phase (e.g., an oil-in-water orwater-in-oil-in-water emulsion) or a continuous oil phase (e.g.,water-in-oil or oil-in-water-in-oil emulsion). The oil phase of theemulsion may include silicone oils, non-silicone oils such ashydrocarbon oils, esters, ethers, and mixtures thereof. The aqueousphase may include water and water-soluble ingredients (e.g.,water-soluble moisturizing agents, conditioning agents, anti-microbials,humectants and/or other skin care actives). In some instances, theaqueous phase may include components other than water, including but notlimited to water-soluble moisturizing agents, conditioning agents,anti-microbials, humectants and/or other water-soluble skin careactives. In some instances, the non-water component of the compositioncomprises a humectant such as glycerin and/or other polyol(s).

In some instances, the compositions herein are in the form of anoil-in-water (“O/W”) emulsion that provides a sensorial feel that islight and non-greasy. Suitable O/W emulsions herein may include acontinuous aqueous phase of more than 50% by weight of the composition,and the remainder being the dispersed oil phase. The aqueous phase mayinclude 1% to 99% water, based on the weight of the aqueous phase, alongwith any water soluble and/or water miscible ingredients. In theseinstances, the dispersed oil phase will typically be present at lessthan 30% by weight of composition (e.g., 1% to 20%, 2% to 15%, 3% to12%, 4% to 10%, or even 5% to 8%) to help avoid some of the undesirablefeel effects of oily compositions. The oil phase may include one or morevolatile and/or non-volatile oils (e.g., botanical oils, silicone oils,and/or hydrocarbon oils). Some nonlimiting examples of oils that may besuitable for use in the present compositions are disclosed in U.S. Pat.No. 9,446,265 and U.S. Publication No. 2015/0196464.

The carrier may contain one or more dermatologically acceptablediluents. As used herein, “diluent” refers to materials in which theskin care actives herein can be dispersed, dissolved, or otherwiseincorporated. Some non-limiting examples of hydrophilic diluents includewater, organic hydrophilic diluents such as lower monovalent alcohols(e.g., C₁-C₄) and low molecular weight glycols and polyols, includingpropylene glycol, polyethylene glycol (e.g., molecular weight of 200 to600 g/mole), polypropylene glycol (e.g., molecular weight of 425 to 2025g/mole), glycerol, butylene glycol, 1,2,4-butanetriol, sorbitol esters,1,2,6-hexanetriol, ethanol, isopropanol, sorbitol esters, butanediol,ether propanol, ethoxylated ethers, propoxylated ethers and combinationsthereof.

Conditioning Agents

The compositions herein may include 0.1% to 50% by weight of aconditioning agent (e.g., 0.5% to 30%, 1% to 20%, or even 2% to 15%).Adding a conditioning agent can help provide the composition withdesirable feel properties (e.g., a silky, lubricious feel uponapplication). Some non-limiting examples of conditioning agents include,hydrocarbon oils and waxes, silicones, fatty acid derivatives,cholesterol, cholesterol derivatives, diglycerides, triglycerides,vegetable oils, vegetable oil derivatives, acetoglyceride esters, alkylesters, alkenyl esters, lanolin, wax esters, beeswax derivatives,sterols and phospholipids, salts, isomers and derivatives thereof, andcombinations thereof. Particularly suitable examples of conditioningagents include volatile or non-volatile silicone fluids such asdimethicone copolyol, dimethylpolysiloxane, diethylpolysiloxane, mixedC1-30 alkyl polysiloxanes, phenyl dimethicone, dimethiconol,dimethicone, dimethiconol, silicone crosspolymers, and combinationsthereof. Dimethicone may be especially suitable, since some consumersassociate the feel properties provided by certain dimethicone fluidswith good moisturization. Other examples of silicone fluids that may besuitable for use as conditioning agents are described in U.S. Pat. No.5,011,681.

Rheology Modifiers

The compositions herein may include 0.1% to 5% of a rheology modifier(e.g., thickening agent) to provide the composition with suitablerheological and skin feels properties. Some non-limiting examples ofthickening agents include crosslinked polyacrylate polymers,polyacrylamide polymers, polysaccharides, gums and mixtures thereof. Ina particularly suitable example, the composition may include asuperabsorbent polymer thickening agent such as sodium polyacrylate,starch grafted sodium polyacrylate, or a combination of these. Somenon-limiting examples of superabsorbent polymer thickeners are describedin, for example, U.S. Pat. No. 9,795,552.

Some consumers find compositions that use silicone fluids asconditioning agents to be undesirably greasy or heavy feeling. Thus, itmay be desirable to provide a composition that is free of orsubstantially free of silicone fluid. It may also be desirable to tailora superabsorbent polymer thickener to provide the composition with alight, airy feel, for example, by adjusting the amount of water in thecomposition, the water:oil ratio (e.g., 12:1 to 1:1), and/or the ratioof water to thickener or oil to thickener.

Emulsifiers

When the dermatologically acceptable carrier is in the form of anemulsion, it may be desirable to include an emulsifier to provide astable composition (e.g., does not phase separate). When included, theemulsifier may be present at an amount of 0.1% to 10% (e.g., 1% to 5%,or 2%-4%). Emulsifiers may be nonionic, anionic or cationic. Somenon-limiting examples of emulsifiers that !nay be suitable for useherein are disclosed in U.S. Pat. Nos. 3,755,560; 4,421,769; andMcCutcheon's Detergents and Emulsifiers, North American Edition, pages317-324 (1986).

Other Optional Ingredients

The present composition may optionally include one or more additionalingredients commonly used in cosmetic compositions (e.g., colorants,skin care actives, anti-inflammatory agents, sunscreen agents,emulsifiers, buffers, rheology modifiers, combinations of these and thelike), provided that the additional ingredients do not undesirably alterthe skin health or appearance benefits provided by the presentcompositions. The additional ingredients, when incorporated into thecomposition, should be suitable for use in contact with human skintissue without undue toxicity, incompatibility, instability, allergicresponse, and the like. Some nonlimiting examples of additional activesinclude vitamins, minerals, peptides and peptide derivatives, sugaramines, sunscreens, oil control agents, particulates, flavonoidcompounds, hair growth regulators, anti-oxidants and/or anti-oxidantprecursors, preservatives, protease inhibitors, tyrosinase inhibitors,anti-inflammatory agents, moisturizing agents, exfoliating agents, skinlightening agents, sunless tanning agents, lubricants, anti-acneactives, anti-cellulite actives, chelating agents, anti-wrinkle actives,anti-atrophy actives, phytosterols and/or plant hormones, N-acyl aminoacid compounds, antimicrobials, and antifungals. Other non-limitingexamples of additional ingredients and/or skin care actives that may besuitable for use herein are described in U.S. Publication Nos.2002/0022040; 2003/0049212; 2004/0175347; 2006/0275237; 2007/0196344;2008/0181956; 2008/0206373; 2010/00092408; 2008/0206373; 2010/0239510;2010/0189669; 2010/0272667; 2011/0262025; 2011/0097286; US2012/0197016;2012/0128683; 2012/0148515; 2012/0156146; and 2013/0022557; and U.S.Pat. Nos. 5,939,082; 5,872,112; 6,492,326; 6,696,049; 6,524,598; and6,174,533.

When including optional ingredients in the compositions herein, it maybe desirable to select ingredients that do not form complexes orotherwise undesirably interact with other ingredients in thecomposition, especially pH sensitive ingredients like niacinamide,salicylates and peptides. When present, the optional ingredients may beincluded at amounts of from 0.0001% to 50%; from 0.001% to 20%; or evenfrom 0.01% to 10% (e.g., 50%, 40%, 30%, 20%, 10%, 5%, 4%, 3%, 2%, 1%,0.5% or 0.1%), by weight of the composition.

Method of Use

The present method includes identifying a target portion of skin wheretreatment is desired and applying a composition comprising an effectiveamount of vitamin B₃ compound, pal-KTTKS [SEQ ID NO: 1], and ac-PPYL[SEQ ID NO: 2], and optionally one or more additional skin care actives,to the target portion of skin. The target portion of skin may be on afacial skin surface such as the forehead, perioral, chin, periorbital,nose, and/or cheek) or another part of the body (e.g., hands, arms,legs, back, chest). The person or target portion of skin in need oftreatment may be one that exhibits a telltale sign of aging skin (e.g.,fine lines, wrinkles, hyperpigmented spots). In some instances, a targetportion of skin may not exhibit a sign of skin aging, but a user maystill wish to treat the portion of skin if it is one that commonlyexposed to higher levels of exogenous stressors (e.g., sun exposed skinsuch as facial skin and arm skin). In this way, the present methods andcompositions may be used prophylactically to help delay skin aging.

The composition may be applied to a target portion of skin and, ifdesired, to the surrounding skin at least once a day, twice a day, or ona more frequent daily basis, during a treatment period. When appliedtwice daily, the first and second applications are separated by at least1 to 12 hours. Typically, the composition is applied in the morningand/or in the evening before bed. The treatment period herein is ideallyof sufficient time for the vitamin B₃ compound, pal-KTTKS [SEQ ID NO:1], and ac-PPYL [SEQ ID NO: 2] to improve the appearance of the skin.The treatment period may last for at least 1 week (e.g., about 2 weeks,4 weeks, 8 weeks, or even 12 weeks). In some instances, the treatmentperiod will extend over multiple months (i.e., 3-12 months). In someinstances, the composition may be applied most days of the week (e.g.,at least 4, 5 or 6 days a week), at least once a day or even twice a dayduring a treatment period of at least 2 weeks, 4 weeks, 8 weeks, or 12weeks.

The step of applying the composition may be accomplished by localizedapplication. In reference to application of the composition, the terms“localized”, “local”, or “locally” mean that the composition isdelivered to the targeted area (e.g., a hyperpigmented spot or portionthereof) while minimizing delivery to skin surfaces where treatment isnot desired. The composition may be applied and lightly massaged into anarea of skin. The form of the composition or the dermatologicallyacceptable carrier should be selected to facilitate localizedapplication. While certain embodiments herein contemplate applying acomposition locally to an area, it will be appreciated that compositionsherein can be applied more generally or broadly to one or more skinsurfaces. In certain embodiments, the compositions herein may be used aspart of a multi-step beauty regimen, wherein the present composition maybe applied before and/or after one or more other compositions.

ARE Assay

This method provides a way to quantitate ARE activation using the ARE-32reporter cell line available from CXR Biosciences. The ARE-32 cell lineis a stable MCF7 cell line containing pGL8x-ARE (8 copies of the rat GSTARE linked to the luciferase gene) and pCDNA3.1, which contains theneomycin selectable marker. A detailed description of the ARE-32 cellline and its development and use can be found in “Generation of a StableAntioxidant Response Element-Driven Reporter Gene Cell Line and Its Useto Show Redox-Dependent Activation of Nrf2 by Cancer ChemotherapeuticAgents.” Cancer Res 2006; 66(22): 10983-94. A general schematic for howan ARE reporter assay operates to identify agents that promotetranscription off the ARE is described in U.S. Publication No.2011/0262570.

Summary of the Method

The ARE Assay uses expanded and cryopreserved passaged stocks. The cellsare expanded over 4-5 days in culture flasks and passaged every 3-4days, when cells are approximately 80% confluent. When cells are >70%confluent or ready to seed into 96 well plates, the cells aretrypsinized, seeded and grown in 96-well plates. After growing for 1 dayin 96-well plates, the media is replaced with fresh treatment media(phenol red free, no FBS) and cells are treated with compounds andincubated overnight (24 hours). Post treatment, cells are rinsed with 1×PBS, lysed and receive the luciferase kit reagent and luminescence ismeasured.

Equipment:

-   -   Biological Safety Cabinet    -   Multi-channel pipette    -   Inverted Microscope    -   Water Bath    -   Bench top centrifuge    -   Incubator    -   Plate Reader (that can read luminescence)    -   Corning 3275 cell culture flask (or comparable)    -   Pipets and pipette controller (ex/Pipet Boy, Drummond Pipet)    -   Aspirator that uses pipet tips and hooks to house vacuum    -   96-well plate (Costar, Cat #3903 or 3610)

Reagents and Materials:

-   -   Dulbecco's Modified Eagle Medium (DMEM) (Gibco, Cat #11054-020)    -   Fetal Bovine Serum One Shot, Heat Inactivated (Gibco, Cat        #A31604)    -   Geneticin G418 sulphate (G418) (Gibco, Cat #10131-027)    -   Penicillin-Streptomycin 100× (Gibco, Cat #15140-148)    -   GlutaMAX Supplement 100× (Gibco 35050-061)    -   0.25% Trypsin EDTA (Gibco 25200-056)    -   0.4% Trypan Blue if using hemocytometer to count cells    -   1× PBS    -   Luciferase Assay System including lysis buffer (Promega, Cat        #E4530)

Plating Media:

-   -   500 ml DMEM (Gibco, Cat #11054-020)    -   0.8 mg/ml G418    -   5 ml GlutaMAX    -   50 ml PBS    -   5 ml Pen/Strep

Treatment Media:

-   -   500 ml DMEM (Gibco, Cat #11054-020)    -   5 ml GlutaMAX    -   5 ml Pen/Strep

Starting New Cultures:

Frozen AREC32 cells (frozen in 90% FBS and 10% DMSO) are quickly thawedand placed in a 50 ml conical tube with 25 ml plating media. Centrifugeat 1500-2000 RPM for 5 minutes. Remove media without disturbing cellpellet. Re-suspend cells in 12 ml media and add to T-75 tissue cultureflask. Cells should be at >80% confluency in 4-5 days and can be split1:3 into T-150 flasks to grow cells for seeding plates if more cells areneeded.

Maintaining and Sub-Culturing Cells:

Cells are maintained and plated in 96-well plates with plating media.Subculture (passage cells) every 3-4 days or when ˜75-80% confluent. Topassage cells, aspirate media from flask and add 6 ml 0.25% trypsin.Tilt flask in all directions to distribute the trypsin over the bottomof the flask and place in incubator. After 2-3 minutes observe cellsunder microscope to see if detached. If cells have not detachedcompletely place in incubator for an additional 2-3 minutes. Oncedetached, add 6 ml maintenance media to neutralize trypsin and pipetinto centrifuge tube. Centrifuge at 1200 RPM for 5 minutes. Remove mediawithout disturbing cell pellet. Re-suspend pellet in 12 ml maintenancemedia and add to flask. If splitting or detaching cells from a T-150flask double all volumes and follow same procedures.

Plating Cells:

Aspirate media from flask and add appropriate volume of 0.25% trypsindepending on size of flask (6 ml for T-75 and 12 ml for T-150 flask).Swirl flask to distribute trypsin on bottom of flask and return toincubator for 2-3 minutes. Observe cells under microscope. If cells havenot detached completely place in incubator for an additional 2-3minutes. Once cells are detached add DMEM equal to the volume oftrypsin, mix gently and place in centrifuge tube. Centrifuge at 12000RPM for 5 minutes. Aspirate media without disturbing cell pellet.Re-suspend pellet in 10 ml DMEM. Dilute the cells 1:5 using 50 μlcells+200 μl DMEM. Use 20 μl of this dilution and 20 μl0.4% trypan blue.Mix and add 10 μl/chamber to disposable hemocytometer and insert intoCountess II Instrument. Non-viable cells will be blue, viable cells willbe unstained. Cell seeding density may vary from ˜10,000 cells/well ormore depending on cell passage growth. Cell density should be approx.70-80% on day of treatment/dosing. On day 2, replace plating media withtreatment media, but do not add the pen/strep. Media must be free ofphenol red.

Positive Control:

Tert-Butylhydroquinone (tBHQ) (Aldrich, Cat #11,294-1). Prepare 100 mMstock (1M=166.21 mg/ml; 100 mM=16.62 mg/ml). Dilute to 750 μM (1:133)using 30 μl of 100 mM tBHQ and 3.62 ml treatment media. 2 μl of positivecontrol should yield a final concentration of 7.5 μM in the test well.

Test Materials:

Test materials may be prepared in DMSO or water, with a finalconcentration of DMSO not to exceed 1%.

Procedure: Cell Preparation and Treatment:

-   -   1. In a 96 well-plate, seed 1.5×10⁴ cells/well in 100 μl plating        media.    -   2. After sitting for 15 minutes at room temperature, place the        cells in the incubator.    -   3. Incubate the cells at 37° C., 5% CO2, and 95% humidity for 24        hrs.    -   4. Replace the media with 99 μl of treatment media and treat        with test compounds (2 μl/well), vehicle control (2 μl/well) and        positive control tBHQ (2 μl/well).    -   5. Add 99 μl of media after treatment, final assay volume 200        μl: Adding half the media after dosing insures better        distribution of the materials.    -   6. Incubate the cells at 37° C., 5% CO2, and 95% humidity for        another 24 hrs.    -   7. Remove media and wash the cells once with 100 μl 1× PBS        buffer. Remove PBS and follow instructions for luciferase assay.

Luciferase Assay:

-   -   1. Prepare 1× lysis reagent by adding 4 volumes of water to 1        volume of 5× lysis reagent.    -   2. Add 20 μl of lysis buffer per well. Gently shake plate to        distribute buffer in well. Place plate in 80° C. freezer for 15        minutes to facilitate lysis. Thaw plate completely and confirm        lysis under microscope.    -   3. Prepare Luciferase Assay Reagent by adding Luciferase Assay        Buffer to the vial of lyophilized Luciferase Assay Substrate        according to the manufacturer's instructions. Mix gently. Add        100 μl Luciferase Assay Reagent per well. Ensure there are no        air bubbles.    -   4. Read plate immediately with plate reader (e.g., a Synergy™        Neo2 brand microplate reader available from BioTek).

Calculation of Data:

ARE activation value=Test sample luminescence/(luminescence of averageof vehicle controls)

Vehicle Control=Average cells+DMSO or water (n=8)

Gene Modulation Assay

This method provides a way to measure the ability of a compound ormaterial to modulate the expression of a target gene.

-   -   Cells: tert keratinocytes (tKC)    -   BJ Fibroblasts    -   Plating: Cells are plated the day before treatment.    -   For tert keratinocytes: 100,000 cells/well in 2 ml volume of        medium/well for 12-well plates (e.g., Collagen I coated plates,        Corning cat #356500), or 50,000 cells/well in 1 ml volume of        medium/well for 24-well plates.    -   For BJ fibroblasts: 88,000 cells/well in 2 ml volume of        medium/well for 12-well plates (e.g., Corning cat #3512), or        44,000 cells/well in 1 ml volume of medium/well for 24-well        plates.    -   Medium: For tert keratinocytes: EpiLife (e.g., Thermo Fisher        Scientific cat #MEPI500CA+HKGS (e.g., Thermo Fisher Scientific        cat #S-001-5).    -   For BJ Fibroblasts: EMEM (e.g., ATCC cat #30-2003)+10% FBS        (e.g., HyClone cat #SH30071.02).

Wafergen Process: Total RNA Purification and qPCR

Cell lysates are thawed at 4° C. and then isolated using the Biomek FxPand the RNAdvance Tissue Isolation kit (Beckman Coulter, p/n A32646).The resulting RNA is quantified using the Nandrop 8000 (Nanodrop,ND-8000). cDNA is generated using 500 ng of Total RNA and AppliedBiosystems High Capacity cDNA with Reverse Transcription kit (AppliedBiosystems p/n 4368814). cDNA, assays, and dilutions of PrimeTimeGeneExpression MasterMix (IDT, p/n 1055771) are plated onto a WafergenMyDesign SmartChip (TakaraBio, p/n 640036) using the WafergenNanodispenser. The chip is then loaded into the SmartChip cycler andqPCR performed using the following PCR conditions:

Hold stage: 50° C. for 2 minutes (warm up), then 95° C. for 10 minutes;

PCR stage (40 cycles): 95° C. for 15 seconds, then 60° C. for 1 minute.

Export data in .txt file format for analysis.

EXAMPLES Example 1: Formulations

Table 1 below provides examples of the present skin care compositions.The exemplary compositions are made by blending the A phase componentswith a suitable mixer (e.g., Tekmar RW20DZM or equivalent) and heatingto a temperature of 70-80° C. and maintaining the temperature whilestirring. Separately, the B phase components are blended with a suitablemixer and heated to 70-75° C., while maintaining temperature duringmixing. Phase B is added to Phase A while mixing well to form anoil-in-water (O/W) emulsion. The emulsion is then milled using asuitable mill (e.g., Tekmar T-25 or equivalent) for 5 minutes. When theemulsion is at 60° C., phase C is added while continuing to mix. At 40°C., the ingredients of phase D and E are added to the emulsion. Theemulsion is then milled for 5 minutes to provide a uniform composition.

TABLE 1 Component I II III IV V VI VII VIII IX Phase A Water qs qs qs qsqs qs qs qs qs Glycerol 5.00 7.00 3.00 15.0 7.00 5.00 5.00 3.00 5.00Disodium EDTA 0.10 0.05 0.10 0.10 0.05 0.05 0.05 0.05 0.10 Phase BDimethicone 5 cSt — — — — — — — 10.0 15.0 Dimethicone and Dimethicone —— — — — — — 13.0 15.0 Crosspolymer Laureth-4 — — — — — — — 0.25 0.35Polysorbate 20 — — — — — — — 0.15 0.25 Tapioca Starch and — — — — — — —2.50 3.50 Polymethylsilsesquioxane Avobenzone — — — 3.00 — 3.00 — — —Homosalate — — — 15.0 — 10.0 — — — Octisalate — — — 5.00 — 5.00 — — —Octocrylene — — — 2.60 — 9.00 — — — Isopropyl Isostearate 5.00 2.50 1.00— — — — — — Isohexadecane 1.00 1.50 3.00 — — — — — — Cetyl Alcohol 0.250.50 0.32 0.40 0.40 0.30 0.50 — — Tocopherol Acetate 0.50 0.25 1.00 0.250.25 0.25 — — PEG-100 Stearate 0.20 0.10 0.10 0.30 0.10 0.20 0.10 — —Stearyl Alcohol 0.50 1.50 0.40 0.60 0.50 0.40 0.60 — — Behenyl Alcohol0.40 1.00 0.50 0.50 0.40 0.35 0.50 — — Ethyl Paraben 0.20 0.15 0.20 0.25— — — — — Propyl Paraben 0.10 0.15 0.10 0.15 — — — — —Polymethylsilsesquioxane 1.25 2.50 1.00 — — — — — — Phase C TitaniumDioxide — 0.50 — 0.25 — — — — — Tapioca Starch and — — — — — 12.0 — —Polymethylsilsesquioxane Vinyl Dimethicone/Methicone 1.50 1.50 3.50 5.00— 7.50 — — Silsesquioxane Crosspolymer Sodium Polyacrylate Starch — — —— 1.50 1.00 1.50 — — Hydroxyethyl acrylate/sodium 2.00 1.50 2.50 2.00 —— — 1.25 2.00 acryloyldimethyltaurate copolymer Phase D Water 5 10 10 510 10 10 5 10 Pal-KTTKS [SEQ ID NO: 1] 0.0001 1 0.5 0.25 0.1 0.05 0.0250.01 0.001 Ac-PPYL [SEQ ID NO: 2] 0.0001 2 0.5 0.5 0.05 0.1 0.0125 0.0050.002 Niacinamide 0.05 1 3.5 2 4 5 10 5 0.5 Dexpanthenol 0.5 0.5 0.5 1 11.5 0.25 1 0.5 Phase E Benzyl alcohol 0.25 0.40 0.25 0.50 — — — — —Hexanediol and Caprylyl Glycol — — — — 0.70 0.80 0.70 0.70 1.00Phenoxyethanol — — — — 0.3 0.4 0.5 0.20 0.25 Dimethicone/dimethiconol0.5 1.00 2.00 1.00 2.00 2.00 1.00 1.75 1.00

Example 2: Vitamin B3 Compound, Pal-KTTKS [SEQ ID NO: 1], and Ac-PPYL[SEQ ID NO: 2] Synergistically Upregulate NRF2 [SEQ ID NO: 3].

This example demonstrates the ability of a combination of niacinamide,pal-KTTKS [SEQ ID NO: 1], and ac-PPYL [SEQ ID NO: 2] to synergisticallyactivate the ARE Test compositions and control compositions wereprepared as described above in the ARE Assay and tested accordingly. Thevitamin B₃ compound used in this example is niacinamide [Sigma cat#N5535], the pal-KTTKS is PROMATRIXYL from Sederma (France), and theac-PPYL is SYNIORAGE from BASF Care Creations (New Jersey). The resultsof the test are summarized below in Table 2. N+P+A refers to thecombination of niacinamide (N), pal-KTTKS (P) [SEQ ID NO: 1], andac-PPYL (A) [SEQ ID NO: 2].

Synergy Factor is calculated as:

$\frac{{Observed}{luminescence}{for}{the}{combination}{of}{ingredients}}{{Sum}{of}{the}{individual}{ingredient}{luminscence}{values}}$

A synergy factor greater than 1.00 with p-value≤0.05 indicates astatistically significant synergistic effect. Preferred synergy factorsare greater than 1.3.

TABLE 2 Synergistic ARE activation Concentration (ppm) Average NetLuminescence Pal- Ac- Pal- Ac- N + P + A N + P + A Synergy N KTTKS PPYLN KTTKS PPYL (observed) (expected) Factor p-value 500 1 2 53 24.3 61200.3 138.3 1.45 0.0011 500 1 1 53 24.3 51 193.3 128.3 1.51 0.0025 500 10.5 53 24.3 26 182.3 103.3 1.77 0.0017 500 1 0.1 53 24.3 22 174.7 99.31.76 0.0009 500 1 0.05 53 24.3 12 106.7 89.3 1.19 0.1568 50 1 2 45 24.364 171.3 133.3 1.29 0.0208 50 1 1 45 24.3 56 184.0 125.3 1.47 0.0027 501 0.5 45 24.3 14.7 172.0 84 2.05 0.0040 50 1 0.1 45 24.3 15.7 145.7 851.71 0.0014 50 1 0.05 45 24.3 19.4 104.0 88.7 1.17 0.1316 10 1 2 12 24.361 138.3 97.3 1.42 0.0028 10 1 1 12 24.3 51 128.7 87.3 1.47 0.0041 10 10.5 12 24.3 26 88.7 62.3 1.42 0.0188 10 1 0.1 12 24.3 22 83.3 58.3 1.430.0054 10 1 0.05 12 24.3 12 63.0 48.3 1.30 0.1095 1 1 2 8 24.3 64 125.096.3 1.30 0.0146 1 1 1 8 24.3 56 116.0 88.3 1.31 0.0398 1 1 0.5 8 24.314.7 71.7 47 1.52 0.0304 1 1 0.1 8 24.3 15.7 63.3 48 1.32 0.0719 1 10.05 8 24.3 20 56.0 52.3 1.07 0.6529

As can be seen in Table 2, the data suggest that ratios of niacinamideto pal-KTTKS [SEQ ID NO: 1] to ac-PPYL [SEQ ID NO: 2] (N:P:A) of between500:1:2 and 1:1:0.5 synergistically activate the ARE. However, ratios ofN:P:A of 500:1:0.05, 50:1:2, 10:1:0.05, 1:1:2, and 1:1:0.05 do notappear to provide a synergy factor of greater than 1.3. Thus, it can beimportant to select the right combination of vitamin B₃ compound,pal-KTTKS [SEQ ID NO: 1] to ac-PPYL, as illustrated in Table 2, toprovide the desired synergistic effect.

Example 3: Tetrapeptide Specificity Needed for Synergy

This example demonstrates the importance of selecting a specifictetrapeptide to provide the desired synergistic activation of the ARE.In this test, the amino acids from ac-PPYL [SEQ ID NO: 2] wererearranged to form a new tetrapeptide, ac-YPLP. Test compositions andcontrol compositions were prepared as described above in the ARE Assayand tested accordingly. The results of the test are summarized in Table3 below.

TABLE 3 ARE activation and Specificity of Ac-PPYL to Ac-YPLPConcentration (ppm) Average Net Luminescence Pal- Ac- Ac- Pal- Ac- Ac-N + P + A N + P + A Synergy N kttks ppyl yplp N kttks ppyl yplp(expected) (observed) Factor p-value 500 1 2 0 45 21 57 123 184 1.500.0018 500 1 0 2 45 21 39 105 96 0.91 0.3476

Surprisingly, as can be seen in Table 3, a tetrapeptide with the sameamino acids as ac-PPYL [SEQ ID NO: 2], but arranged in a differentorder, does not provide the desired synergistic effect. These datasuggest that the specific peptide sequence is important for providingthe desired synergy.

Example 4: Synergistic Upregulation of Genes Involved in in CellularAnti-Oxidancy and Repair

This example demonstrates the ability of a combination of niacinamide,pal-KTTKS [SEQ ID NO: 1] and ac-PPYL [SEQ ID NO: 2] to synergisticallyupregulate SLFNS [ SEQ ID NO: 4], GDE1 [ SEQ ID NO: 5], MINPP1 [ SEQ IDNO: 6] and HMGCL [ SEQ ID NO: 7], which are involved in cellularanti-oxidancy and repair processes. Test compositions and controlcompositions were prepared as described above in the Gene ModulationAssay and tested accordingly. The pal-KTTKS used in this example isPROMATRIXYL brand pal-KTTKS from Sederma (France), and the ac-PPYL isSYNIORAGE brand tetrapeptide from BASF Care Creations (New Jersey). Theresults of the test are summarized below in Table 4. Fold change shownin Table 4 is based on the combination (N+P+A) versus the sum ofindividual treatments (N, P and A). A p-value of 0.05 or less isconsidered significant.

TABLE 4 Synergistic Upregulation of Selected Biomarkers NiacinamidePal-KTTKS Ac-PPYL Fold (ppm) (ppm) (ppm) Biomarker Change p-value 500 11 GDE1 1.4228 0.0198 500 1 1 HMGCL 1.4687 0.0368 500 1 1 MINPP1 1.35720.0195 500 1 1 SLFN5 1.6198 0.0011

Example Combinations

-   -   A. A skin care composition, comprising:        -   1) a combination of a vitamin B3 compound, palmitoyl            pentapeptide-4 (pal-KTTKS) [SEQ ID NO: 1], and acetyl            tetrapeptide-11 (ac-PPYL) [SEQ ID NO: 2], wherein the            combination of vitamin B3 compound, pal-KTTKS and ac-PPYL            increases activation of a cell's Antioxidant Response            Element (ARE) according to the ARE Assay; and        -   2) a dermatologically acceptable carrier.    -   B. The skin care composition of paragraph A, wherein the        combination of vitamin B3 compound, pal-KTTKS and ac-PPYL        synergistically boosts activation of the ARE.    -   C. The composition of paragraph A or B, wherein the combination        of vitamin B3 compound, pal-KTTKS [SEQ ID NO: 1], and ac-PPYL        [SEQ ID NO: 2] exhibit a synergy factor of at least 1.3.    -   D. The composition any preceding paragraph, wherein the        combination of vitamin B3 compound, pal-KTTKS [SEQ ID NO: 1],        and ac-PPYL [SEQ ID NO: 2] synergistically upregulates at least        one gene selected from the group consisting of Nuclear Factor        E2-Related Factor 2 (NRF2) [SEQ ID NO: 3], Schlafen Family        Member 5 (SLFNS) [SEQ ID NO: 4], Glycerophosphodiester        Phosphodiesterase 1 (GDE1) [SEQ ID NO: 5], Multiple        Inositol-Polyphosphate Phosphatase 1 (MINPP1) [SEQ ID NO: 6],        and 3-Hydroxy-3-Methylglutaryl-CoA Lyase (HMGCL) [SEQ ID NO: 7].    -   E. The composition of any preceding paragraph, wherein the        vitamin B3 compound is present at 0.05% to 10% by weight of the        composition.    -   F. The composition of any preceding paragraph, wherein the        vitamin B3 compound is selected from the group consisting of        niacinamide, nicotinic acid, nicotinyl alcohol, and combinations        thereof.    -   G. The composition of paragraph F, wherein the vitamin B3        compound is niacinamide.    -   H. The composition of any preceding paragraph, wherein the        pal-KTTKS [SEQ ID NO: 1] is present at 0.0001% to 2% by weight        of the composition.    -   I. The composition of any preceding paragraph, wherein the        ac-PPYL [SEQ ID NO: 2] is present at 0.0001% to 2% by weight of        the composition.    -   J. The composition of any preceding paragraph, wherein a ratio        of vitamin B3 compound to pal-KTTKS [SEQ ID NO: 1] to ac-PPYL        [SEQ ID NO: 2] is between 500:1:2 and 1:1:0.5.    -   K. The composition of any preceding paragraph, further        comprising at least one additional ingredient selected from        vitamins, minerals, peptides, sugar amines, sunscreen agents,        oil control agents, flavonoid compounds, anti-oxidants, protease        inhibitors, tyrosinase inhibitors, anti-inflammatory agents,        moisturizing agents, exfoliating agents, skin lightening agents,        anti-acne agents, anti-wrinkle agents, phytosterols, N-acyl        amino acid compounds, antimicrobials, antifungals, pH adjustors,        thickening agents, preservatives, and combinations thereof.    -   L. A method of treating oxidative stress in skin, comprising:    -   identifying a target portion of skin where treatment is desired;        and    -   applying the skin care composition of any preceding paragraph        thereto.    -   M. The method of paragraph L, wherein the composition improves        the appearance of a visible sign of skin aging.    -   III. A method of upregulating NRF2 [SEQ ID NO: 3] in a skin        cell, comprising: contacting a skin cell with an effective        amount of a vitamin B3 compound, pal-KTTKS [SEQ ID NO: 1], and        ac-PPYL [SEQ ID NO: 2] in combination, wherein the effective        amount of vitamin B3 compound, pal-KTTKS [SEQ ID NO: 1], and        ac-PPYL [SEQ ID NO: 2] synergistically increases activation of        the Antioxidant Response Element according the ARE Assay.

The dimensions and values disclosed herein are not to be understood asbeing strictly limited to the exact numerical values recited. Instead,unless otherwise specified, each such dimension is intended to mean boththe recited value and a functionally equivalent range surrounding thatvalue. For example, a dimension disclosed as “40 mm” is intended to mean“about 40 mm”.

Every document cited herein, including any cross referenced or relatedpatent or application and any patent application or patent to which thisapplication claims priority or benefit thereof, is hereby incorporatedherein by reference in its entirety unless expressly excluded orotherwise limited. The citation of any document is not an admission thatit is prior art with respect to any invention disclosed or claimedherein or that it alone, or in any combination with any other referenceor references, teaches, suggests or discloses any such invention.Further, to the extent that any meaning or definition of a term in thisdocument conflicts with any meaning or definition of the same term in adocument incorporated by reference, the meaning or definition assignedto that term in this document shall govern.

While particular embodiments of the present invention have beenillustrated and described, it would be obvious to those skilled in theart that various other changes and modifications can be made withoutdeparting from the spirit and scope of the invention. It is thereforeintended to cover in the appended claims all such changes andmodifications that are within the scope of this invention.

What is claimed is:
 1. A skin care composition, comprising: a) acombination of a vitamin B₃ compound, palmitoyl pentapeptide-4(pal-KTTKS) [SEQ ID NO: 1], and acetyl tetrapeptide-11 (ac-PPYL) [SEQ IDNO: 2], wherein the combination of vitamin B₃ compound, pal-KTTKS andac-PPYL synergistically increases activation of a cell's AntioxidantResponse Element (ARE) according to the ARE Assay and exhibit a synergyfactor of at least 1.3; and b) a dermatologically acceptable carrier; c)at least one additional ingredient chosen from vitamins, minerals,peptides, sugar amines, sunscreen agents, oil control agents, flavonoidcompounds, anti-oxidants, protease inhibitors, tyrosinase inhibitors,anti-inflammatory agents, moisturizing agents, exfoliating agents, skinlightening agents, anti-acne agents, anti-wrinkle agents, phytosterols,N-acyl amino acid compounds, antimicrobials, antifungals, pH adjustors,thickening agents, preservatives, or mixtures thereof; wherein a ratioof the vitamin B3 compound to the pal-KTTKS [SEQ ID NO: 1] and to theac-PPYL [SEQ ID NO: 2] is between 500:1:2 and 1:1:0.5
 2. The compositionof claim 1, wherein the combination of vitamin B₃ compound, pal-KTTKS[SEQ ID NO: 1], and ac-PPYL [SEQ ID NO: 2] synergistically upregulatesat least one gene selected from the group consisting of Nuclear FactorE2-Related Factor 2 (NRF2) [SEQ ID NO: [3], Schlafen Family Member 5(SLFNS) [SEQ ID NO: [41, Glycerophosphodiester Phosphodiesterase 1(GDE1) (SEQ ID NO: [5], Multiple Inositol-Polyphosphate Phosphatase 1(MINPP1) [SEQ ID NO: [6], and 3-Hydroxy-3-Methylglutaryl-CoA Lyase(HMGCL) [SEQ ID NO: [7].
 3. The composition of claim 1, wherein thevitamin B₃ compound is present at about 0.05% to about 10% by weight ofthe composition.
 4. The composition of claim 1, wherein the vitamin B₃compound is selected from the group consisting of niacinamide, nicotinicacid, nicotinyl alcohol, and combinations thereof.
 5. The composition ofclaim 4, wherein the vitamin B₃ compound is niacinamide.
 6. Thecomposition of claim 1, wherein the pal-KTTKS [SEQ ID NO: 1] is presentat about to about 2% by weight of the composition.
 7. The composition ofclaim 1, wherein the ac-PPYL [SEQ ID NO: 2] is present at about to about2% by weight of the composition.
 8. A method of treating oxidativestress in skin: a) identifying a target portion of skin where treatmentis desired; and b) applying the skin care composition of claim 1 to thetarget portion of skin during a treatment period; d) wherein theeffective amount of vitamin B₃ compound, pal-KTTKS [SEQ ID NO: 1] andac-PPYL [SEQ ID NO: 2] is sufficient to increase activation of theAntioxidant Response Element (ARE) according to the ARE Assay.
 9. Themethod of claim 8, wherein the method improves the appearance of avisible sign of skin aging.